Somatostatin antagonists

ABSTRACT

The invention features somatostatin antagonists having a D-amino acid at the second residue.

CROSS REFERENCE TO RELATED APPLICATION

This is a divisional application of U.S. application Ser. No.09/670,249, filed Sep. 26, 2000 now U.S. Pat. No. 6,703,481, which is acontinuation of application Ser. No. 08/855,204, filed May 13, 1997 nowU.S. Pat. No. 6,626,229, a non-provisional application that claims thebenefit of U.S. application Ser. No. 60/032,358, filed Dec. 4, 1996,each of which is hereby incorporated herein by reference in itsentirety.

BACKGROUND OF THE INVENTION

Native somatostatin is comprised of both a 14-amino acid isoform(somatostatin-14) and a 28-amino acid isoform (somatostatin-28). Heiman,et al., Neuroendocrinology, 45:429-436 (1987). Because of the shorthalf-life of the native somatostatin, various somatostatin analogs havebeen developed, e.g., for the treatment of acromegaly. Raynor, et al.,Molecular Pharmacol. 43:838 (1993). Five distinct somatostatin receptorshave been identified and characterized. Hoyer, et al.,Naunyn-Schmiedeberg's Arch. Pharmacol., 350:441 (1994). Somatostatinproduces a variety of effects, including modulation of hormone release,e.g., growth hormone, glucagon, 15-insulin, amylin, and neurotransmitterrelease. Some of these effects have been associated with its binding toa specific somatostatin receptor. For example, the inhibition of growthhormone has been attributed to the somatostatin type-2 receptor(“SSTR-2”) (Raynor, et al., Molecular Pharmacol. 43:838 (1993); Lloyd,et al., Am. J. Physiol. 268:G102 (1995)) while the inhibition of insulinhas been attributed to the somatostatin type-5 receptor (“SSTR-5”) (Coy,et al. 197:366-371 (1993)). The following invention relates to a novelclass of somatostatin analogs which are antagonists to somatostatinreceptors.

SUMMARY OF THE INVENTION

The invention features a compound of the formula:

wherein

A¹ is a D- or L-isomer of an aromatic amino acid, or is deleted;

A² is a D-isomer selected from the group consisting of Cys, Pen, anaromatic amino acid, or an aliphatic amino acid;

A³ is an aromatic amino acid;

A⁴ is Trp or D-Trp;

A⁶ is Thr, Thr(Bzl), Gly, Ser, an Eaa, or an aliphatic amino acid;

A⁷ is Cys, Pen, or an aromatjkic or an aliphatic amino acid;

A⁸ is a D- or L-isomer selected from the group consisting of Thr, Ser,an aromatic amino acid, or an, aliphatic amino acid;

each of R₁ and R₂, is, independently, H or substituted (e.g., one tofour times) or unsubstituted lower alkyl, aryl, aryl lower alkyl,heterocycle, heterocycle lower alkyl, E₁SO₂ or E₁CO (where E₁ is aryl,aryl lower alkyl, heterocycle, or heterocycle lower alkyl), where saidsubstituent is halo, lower alkyl, hydroxy, halo lower alkyl, or hydroxylower alkyl; and

R₃ is OH, NH₂, C₁₋₁₂ alkoxy, or NH—Y—CH₂-Z, wherein Y is a C₁₋₁₂hydrocarbon moiety and Z is H, OH, CO₂H, or CONH₂, or R₃, together withthe carbonyl group of A⁸ attached thereto, are reduced to form H, loweralkyl, or hydroxy lower alkyl; provided if A² is D-Cys or D-Pen, and A⁷is Cys or Pen, then a disulfide bond links the sidechains of A² and A⁷,and if A¹ is D-Phe or p-NO₂-Phe; A² is D-Cys; A³ is Phe or Tyr; A⁶ isThr or Val; and A⁷ is Cys; then A⁸ is β-Nal.

In one embodiment, A² is D-Cys, A⁷ is Cys, and A⁴ is D-Trp. In a furtherembodiment, A¹ is an L-aromatic amino acid. In still a furtherembodiment, A¹ and A³, independently, is β-Nal, o-X-Phe (where X is H,OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂), p-X-Phe (where X is H, OH, CH₃,halo, OCH₃, NH₂, CN, or NO₂), m-X-Phe (where X is H, OH, CH₃, halo,OCH₃, NH₂, CN, or NO₂), F₅-Phe, Trp, Dip, 2-Pal, Tyr(Bzl), His, Igl,Tyr(I), Bta, Bip, Npa, or Pal; A⁶ is Thr, Ser, Tle, Thr(Bzl), Abu, Ala,Ile, Leu, Gly, Nle, β-Ala, Gaba, or Val; and A⁸ is the D- or L-isomer ofThr, Dip, F₅-Phe, p-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN,or NO₂), o-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂),m-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂), Igl,Tyr(Bzl), or β-Nal. In yet still another embodiment, A¹ is β-Nal, Npa,Igl, Phe, p-F-Phe, Trp, p-Cl-Phe, or p-CN-Phe; A³ is Tyr, Tyr(I), orPal; A⁶ is Val, Tle, Nle, Ile, or Leu; A⁸ is p-F-Phe, β-Nal, Tyr, Dip,p-Cl-Phe, Igl, or p-CN-Phe; R₁ is H, CH₃CO,4-(2-hydroxyethyl)-1-piperazinylacetyl, or4-(2-hydroxyethyl)-1-piperizineethanesulfonyl; R₂ is H; and R₃ is NH₂.

In another further embodiment, A¹ is a D-aromatic amino acid. In stillanother further embodiment, A¹ is D-β-Nal, D-o-X-Phe (where X is H, OH,CH₃, halo, OCH₃, NH₂, CN, or NO₂), D-p-X-Phe (where X is H, OH, CH₃,halo, OCH₃, NH₂, CN, or NO₂), D-m-X-Phe (where X is H, OH, CH₃, halo,OCH₃, NH₂, CN, or NO₂), D-F₅-Phe, D-Trp, D-Dip, D-2-Pal,D-Tyr(Bzl)-D-His, D-Igl, D-Tyr(I), D-Bta, D-Bip, D-Npa, or D-Pal; A³ isβ-Nal, o-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂),p-X-Phe (where-X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂), m-X-Phe(where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂), F₅-Phe, Trp, Dip,2-Pal, Tyr(Bzl), His, Igl, Tyr(I), Bta, Bip, Npa, or Pal; A⁶ is Thr,Ser, Tle, Thr(Bzl), Abu, Ala, Ile, Leu, Gly, Nle, β-Ala, Gaba, or Val;and A⁸ is the D- or L-isomer of Thr, Dip, F₅-Phe, p-X-Phe (where X is H,OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂), o-X-Phe (where X is H, OH, CH₃,halo, OCH₃, NH₂, CN, or NO₂), m-X-Phe (where X is H, OH, CH₃, halo,OCH₃, NH₂, CN, or NO₂), Tyr(Bzl), Igl, or β-Nal. In yet still a furtherembodiment, A¹ is D-β-Nal, D-Npa, D-Igl, D-Phe, D-p-F-Phe, D-Trp,D-p-Cl-Phe, or D-p-CN-Phe; A³ is Tyr, Tyr(I), or Pal; A⁶ is Val, Tle,Nle, Ile, or Leu; A⁸ is p-F-Phe, β-Nal, Tyr, Dip, p-Cl-Phe, Igl, orp-CN-Phe; R₁ is H, CH₃CO, 4-(2-hydroxyethyl)-1-piperazinylacetyl, or4-(2-hydroxyethyl)-1-piperizineethanesulfonyl; R₂ is H; and R₃ is NH₂.

In still another further embodiment, A¹ is deleted, R¹ is substituted orunsubstituted E₁CO, and R₂ is H. In still a further embodiment, R₁ issubstituted or unsubstituted E₁CO (where E₁ is phenyl, β-naphthylmethyl,β-pyridinylmethyl, or 3-indolylmethyl); A³ is β-Nal, o-X-Phe (where X isH, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂), p-X-Phe (where X is H, OH,CH₃, halo, OCH₃, NH₂, CN, or NO₂), m-X-Phe (where X is H, OH, CH₃, halo,OCH₃, NH₂, CN, or NO₂), F₅-Phe, Trp, Dip, 2-Pal, Tyr(Bzl), His, Igl,Tyr(I), Bta, Bip, Npa, or Pal; A⁶ is Thr, Ser, Tle, Thr(Bzl), Abu, Ala,Ile, Leu, Gly, Nle, β-Ala, Gaba, or Val; and A⁸ is the D- or L-isomer ofThr, Dip, F₅-Phe, p-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN,or NO₂), o-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂),m-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂), Igl,Tyr(Bzl), or β-Nal. In yet still a further embodiment, R₁ is E₁CO (whereE₁ is 4-hydroxy-phenyl, β-naphthylmethyl, or phenyl); A³ is Tyr, Tyr(I),or Pal; A⁶ is Val, Tle, Nle, Ile, or Leu; A⁸ is p-F-Phe, β-Nal, Tyr,Dip, p-Cl-Phe, Igl, or p-CN-Phe; R₃ is NH₂.

In yet still a further embodiment, R₃, together with the carbonyl groupof A⁸ attached thereto, are reduced to form H lower alkyl, or hydroxylower alkyl. In still another further embodiment, A¹ is the D- orL-isomer of β-Nal, o-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN,or NO₂), p-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂),m-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂), F₅-Phe,Trp, Dip, 2-Pal, Tyr(Bzl), His, Igl, Tyr(I), Bta, Bip, Npa, or Pal; A³is β-Nal, o-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂),p-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂), m-X-Phe(where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂), F₅-Phe, Trp, Dip,2-Pal, Tyr(Bzl), His, Igl, Tyr(I), Bta, Bip, Npa, or Pal; A⁶ is Thr,Ser, Tle, Thr(Bzl), Abu, Ala, Ile, Leu, Gly, Nle, β-Ala, Gaba, or Val;and A⁸ is the D- or L-isomer of Thr, Dip, F₅-Phe, p-X-Phe (where X is H,OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂), o-X-Phe (where X is H, OH, CH₃,halo, OCH₃, NH₂, CN, or NO₂), m-X-Phe (where X is H, OH, CH₃, halo,OCH₃, NH₂, CN, or NO₂), Igl, Tyr(Bzl), or β-Nal. In yet still anotherfurther embodiment, A¹ is the D- or L-isomer of β-Nal, Phe, p-F-Phe,Trp, p-Cl-Phe, or p-CN-Phe; A³ is Tyr, Tyr(I), or Pal; A⁶ is Val, Tle,Nle, Ile, or Leu; A⁸ is p-F-Phe, β-Nal, Tyr, Dip, p-Cl-Phe, Igl, orp-CN-Phe; R₁ is H, CH₃CO, 4-(2-hydroxyethyl)-1-piperazinylacetyl, or4-(2-hydroxyethyl)-1-piperizineethanesulfonyl; R₂ is H, and R₃, togetherwith the carboxy group of A⁸ attached thereto, are reduced to form H orCH₃OH.

In another embodiment, A² is a D-aromatic amino acid or a D-aliphaticamino acid, A⁷ is an aromatic amino acid or an aliphatic amino acid, andA⁴ is D-Trp. In a further embodiment, A¹ is an L-amino acid and A² is aD-aromatic amino acid. In still a further embodiment, A¹, A³, and A⁷independently, is β-Nal, o-X-Phe (where X is H, OH, CH₃, halo, OCH₃,NH₂, CN, or NO₂), p-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN,or NO₂), m-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂),F₅-Phe, Trp, Dip, 2-Pal, Tyr(Bzl), His, Igl, Tyr(I), Bta, Bip, Npa, orPal; A² is D-β-Nal, D-o-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂CN, or NO₂) D-p-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, orNO₂), D-m-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂),D-F₅-Phe, D-Trp, D-Dip, D-2-Pal, D-Tyr(Bzl), D-His, D-Igl, D-Tyr(I),D-Bta, D-Bip, D-Npa, or D-Pal; A⁶ is Thr, Ser, Tle, Thr(Bzl), Abu, Ala,Ile, Leu, Gly, Nle, β-Ala, Gaba, or Val; and A⁸ is the D- or L-isomer ofThr, Dip, F₅-Phe, p-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN,or NO₂), o-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂),m-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂), Tyr(Bzl),Igl, or β-Nal. In yet still a further embodiment, A¹ is β-Nal or Phe, A²is D-Cpa or D-Phe; A³ is Phe or Tyr; A⁶ is Abu, Thr, or Val; A⁷ is Phe;and A⁸ is Thr; R₁ is H, CH₃CO, 4-(2-hydroxyethyl)-1-piperazinylacetyl,or 4-(2-hydroxyethyl)-1-piperizineethanesulfonyl; R₂ is H; and R₃ isNH₂.

In another further embodiment, A¹ is a D-amino acid and A² is aD-aromatic amino acid. In still a further embodiment, A¹ and A²,independently, is D-β-Nal, D-o-X-Phe (where X is H, OH, CH₃, halo, OCH₃,NH₂, CN, or NO₂), D-p-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN,or NO₂), D-m-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂),D-F₅-Phe, D-Trp, D-Dip, D-2-Pal, D-Tyr(Bzl), D-His, D-Igl, D-Tyr(I),D-Bta, D-Bip, D-Npa, or D-Pal; A³ and A⁷, independently, is β-Nal,o-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂, CN or NO₂), p-X-Phe(where X is H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂), m-X-Phe (where Xis H, OH, CH₃, halo, OCH₃, NH₂, CN, or NO₂), F₅-Phe, Trp, Dip, 2-Pal,His, Igl, Tyr(I), Bta, Bip, Npa, Tyr(Bzl), or Pal; A⁶ is Thr, Ser, Tle,Thr(Bzl), Abu, Ile, Ile, Leu, Gly, Nle, β-Ala, Gaba, or Val; and A⁸ isthe D- or L-isomer of Thr, Dip, F₅-Phe, p-X-Phe (where X is H, OH, CH₃,halo, OCH₃, NH₂, CN, or NO₂), o-X-Phe (where X is H, OH, CH₃, halo,OCH₃, NH₂, CN, or NO₂), m-X-Phe (where X is H, OH, CH₃, halo, OCH₃, NH₂,CN, or NO₂), Igl, Tyr(Bzl), or β-Nal. In yet still a further embodiment,A¹ is D-β-Nal or D-Phe; A² is D-Cpa or D-Phe; A³ is Phe or Tyr; A⁶ isThr or Val; A⁷ is Phe; and A⁸ is Thr; R₁ is H, CH₃CO,4-(2-hydroxyethyl)-1-piperazinylacetyl, or4-(2-hydroxyethyl)-1-piperizineethanesulfonyl; R₂ is H; and R₃ is NH₂.

Examples of compounds of the present invention include the following:

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂ (Analog No. 2);

(H)(CH₃CO)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂ (Analog No. 5);

(H)-(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

(H)-(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂ (Analog No. 3);

(H)(CH₃CO)-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

(H)-(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

(H)-(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH₂;

(H)(CH₃CO)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH₂;

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-Thr-NH₂;

(H)(CH₃CO)-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-Thr-NH₂;

H₂-Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

(H)(CH₃CO) Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

H₂-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂ (Analog No. 4);

(H)(CH₃CO)Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

H₂-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-Thr-NH₂;

(H)(CH₃CO)-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-Thr-NH₂;

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(CH₃CO)-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(CH₃CO)-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-Thr-NH₂;

H(CH₃CO)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-Thr-NH₂;

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-Thr-NH₂;

(H)(CH₃CO)-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-Thr-NH₂;

H₂-Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(CH₃CO)Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

H₂-Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(CH₃CO)Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

H₂-Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-Thr-NH₂;

(H)(CH₃CO)Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-Thr-H₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-Thr-NH₂;

H₂-Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-Thr-NH₂ (Analog No. 6);

(H)(CH₃CO)-Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-Thr-NH₂;

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-β-Nal-NH₂;

H₂-Phe-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-β-Nal-NH₂;

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Abu-Cys-β-Nal-NH₂;

H₂-Phe-D-Cys-Pal-D-Trp-Lys-Abu-Cys-β-Nal-NH₂;

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-Thr-NH₂;

H₂-Phe-D-Pen-Tyr-D-Trp-Lys-Val-Pen-β-Nal-NH₂; or

H₂-Phe-D-Pen-Pal-D-Trp-Lys-Thr-Pen-Thr-NH₂;

H₂-Dip-D-Cys-Pal-D-Trp-Lys-Val-Cys-Dip-NH₂ (Analog No. 10);

H₂-F₅-Phe-D-Cys-His-D-Trp-Lys-Val-Cys-F₅-Phe-NH₂ (Analog No. 11);

H₂-Dip-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂ (Analog No. 13);

H₂-m-F-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-m-F-Phe-NH₂ (Analog No. 14);

H₂-o-F-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-o-F-Phe-NH₂ (Analog No. 15);

H₂-p-F-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-p-F-Phe-NH₂ (Analog No. 12);

H₂-F₅-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-F₅-Phe-NH₂ (Analog No. 16);

H₂-F₅-Phe-D-Cys-2-Pal-D-Trp-Lys-Val-Cys-F₅-Phe-NH₂ (Analog No. 17);

H₂-β-Nal-D-Cys-His-D-Trp-Lys-Val-Cys-D-Dip-NH₂ (Analog No. 19);

H₂-Dip-D-Cys-His-D-Trp-Lys-Val-Cys-β-Nal-NH₂ (Analog No. 20);

H₂-Dip-D-Cys-His-D-Trp-Lys-Val-Cys-Dip-NH₂ (Analog No. 21);

is 5H₂-β-Nal-D-Cys-His-D-Trp-Lys-Val-Cys-β-Nal-NH₂ (Analog No. 22);

H₂-Trp-D-Cys-Tyr-D-Trp-Lys-Val-Cys-D-β-Nal-NH₂ (Analog No. 24);

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-D-β-Nal-NH₂ (Analog No. 25);

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-D-p-F-Phe-NH₂ (Analog No. 28);

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Tle-Cys-β-Nal-NH₂ (Analog No. 29);

H₂-p-F-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂ (Analog No. 30);

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Nle-Cys-β-Nal-NH₂ (Analog No. 31);

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Ile-Cys-β-Nal-NH₂ (Analog No. 32);

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Gly-Cys-β-Nal-NH₂ (Analog No. 33);

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Ala-Cys-β-Nal-NH₂ (Analog No. 34);

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Leu-Cys-β-Nal-NH₂ (Analog No. 35);

H₂-Bip-D-Cys-Tyr-D-Trp-Lys-Ile-Cys-Bip-NH₂ (Analog No. 36);

H₂-p-F-Phe-D-Cys-His-D-Trp-Lys-Val-Cys-p-F-Phe-NH₂ (Analog No. 38);

H₂-Npa-D-Cys-Pal-D-Trp-Lys-Val-Cys-Tyr-NH₂ (Analog No. 39);

H₂-m-F-Phe-D-Cys-His-D-Trp-Lys-Val-Cys-m-F-Phe-NH₂ (Analog No. 40);

H₂-o-F-Phe-D-Cys-His-D-Trp-Lys-Val-Cys-o-F-Phe-NH₂ (Analog No. 41);

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-Dip-NH₂ (Analog No. 42);

H₂-Cpa-D-Cys-Pal-D-Trp-Lys-Val-Cys-Cpa-NH₂ (Analog No. 43);

H₂-Igl-D-Cys-Pal-D-Trp-Lys-Val-Cys-Igl-NH₂ (Analog No. 44);

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-D-Dip-NH₂ (Analog No. 45);

H₂-β-Nal-D-Cys-3-I-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂ (Analog No. 46);

H₂-p-CN-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-p-CN-Phe-NH (Analog No. 47);

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-D-Dip-NH₂ (Analog No. 48);

H₂-β-Nal-D-Cys-Bta-D-Trp-Lys-Val-Cys-β-Nal-NH₂ (Analog No. 49);

H₂-p-F-Phe-D-Cys-Pal-D-Trp-Lys-Tle-Cys-β-Nal-NH₂ (Analog No. 50);

H₂-Bpa-D-Cys-Pal-D-Trp-Lys-Val-Cys-Bpa-NH₂ (Analog No.

52);

H₂-Iph-D-Cys-Pal-D-Trp-Lys-Val-Cys-Iph-NH₂ (Analog No. 53);

H₂-Trp-D-Cys-Pal-D-Trp-Lys-Tle-Cys-β-Nal-NH₂ (Analog No. 54);

H₂-p-Cl-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂ (Analog No. 55);

H₂-p-Cl-Phe-D-Cys-Pal-D-Trp-Lys-Tle-Cys-β-Nal-NH₂ (Analog No. 56);

H₂-p-Cl-Phe-D-Cys-Pal-D-Trp-Lys-Tle-Cys-p-Cl-Phe-NH₂ (Analog No. 57);

H₂-p-Cl-Phe-D-Cys-Pal-D-Trp-Lys-Cha-Cys-p-Cl-Phe-NH₂;

H₂-p-Cl-Phe-D-Cys-Tyr(I)-D-Trp-Lys-Val-Cys-p-Cl-Phe-NH₂;

H₂-p-Cl-Phe-D-Cys-Tyr(I)-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

H₂-p-Cl-Phe-D-Cys-Tyr(I)-D-Trp-Lys-Tle-Cys-β-Nal-NH₂;

H₂-p-F-Phe-D-Cys-Tyr(I)-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

H₂-p-F-Phe-D-Cys-Tyr(I)-D-Trp-Lys-Tle-Cys-β-Nal-NH₂;

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-β-Nal-NH₂;

(H)(CH₃CO)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-β-Nal-NH₂;

H₂-p-NO₂-Phe-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-β-Nal-NH₂;

(H)(CH₃CO)-p-NO₂-Phe-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-β-Nal-NH₂;

H₂-p-NO₂-Phe-D-Cys-Tyr(Bzl)-D-Trp-Lys-Thr(Bzl)-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-p-NO₂-Phe-D-Cys-Tyr(Bzl)-D-Trp-Lys-Thr(Bzl)-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-p-NO₂-Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-Tyr-NH₂;

H₂-p-NO₂-Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-p-NO₂-Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

H₂-β-Nal-D-Cys-Tyr(Bzl)-D-Trp-Lys-Thr(Bzl)-Cys-β-Nal-NH₂;

(H)(4-2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Tyr(Bzl)-D-Trp-Lys-Thr(Bzl)-Cys-Tyr(Bzl)-NH₂;

H₂-D-Phe-D-Pen-Tyr-D-Trp-Lys-Val-Cys-Thr-NH₂;

H₂-D-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH₂;

H₂-D-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂ (Analog No. 9);

H₂-D-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

H₂-D-Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-Thr-NH₂;

H₂-D-Phe-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-Thr-NH₂;

H₂-D-β-Nal-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-Thr-NH₂;

H₂-D-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-D-β-Nal-NH₂ (Analog No. 26);

H₂-D-p-F-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-D-p-F-Phe-NH₂ (Analog No. 27);

H₂-D-Bip-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂ (Analog No. 37);

H₂-D-Dip-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂ (Analog No. 18);

H₂-D-p-F-Phe-D-Cys-Pal-D-Trp-Lys-Tle-Cys-β-Nal-NH₂ (Analog No. 51);

H₂-D-p-Cl-Phe-D-Cys-Pal-D-Trp-Lys-Tle-Cys-p-Cl-Phe-NH₂ (Analog No. 7);

p-NO₂-D-Phe-D-Cys-Pal-D-Trp-Lys-Thr(Bzl)-Cys-Tyr(Bzl)-NO_(2;)

p-NO₂-D-Phe-D-Cys-Tyr (Bzl)-D-Trp-Lys-Val-Cys-Tyr(Bzl)-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-p-NO₂-D-Phe-D-Cys-Pal-D-Trp-Lys-Thr(Bzl)-Cys-Tyr(Bzl)-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-p-NO₂-D-Phe-D-Cys-Tyr(Bzl)-D-Trp-Lys-Val-Cys-Tyr(Bzl)-NH₂;

(H)(3-phenylpropionyl)-D-Cys-Tyr-D-Trp-Lys-val-Cys-β-Nal-NH₂;

(H)(3-phenylpropionyl)-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

(H)(3-phenylpropionyl)-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(3-phenylpropionyl)-D-Cys-Pal-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(3-phenylpropionyl)-D-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH₂;

(H)(3-phenylpropionyl)-D-Cys-Pal-D-Trp-Lys-Val-Cys-Thr-NH₂;

(H)(3-phenylpropionyl)-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-Thr-NH₂;

(H)(3-phenylpropionyl)-D-Cys-Pal-D-Trp-Lys-Thr-Cys-Thr-NH₂;

(H)(3-[2-naphthyl]propionyl)-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

(H)(3-[2-naphthyl]propionyl)-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

(H)(3-[2-naphthyl]propionyl)-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-β-Nal-NH₂;

(H)(3-[2-naphthyl]propionyl)-D-Cys-Pal-D-Trp-Lys-ahr-Cys-β-Nal-NH₂;

(H)(3-[2-naphthyl]propionyl)-D-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH₂;

(H)(3-[2-naphthyl]propionyl)-D-Cys-Pal-D-Trp-Lys-Val-Cys-Thr-NH₂;

(H)(3-[2-naphthyl]propionyl)-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-Thr-NH₂;

(H)(3-[2-naphthyl]propionyl)-D-Cys-Pal-D-Trp-Lys-Thr-Cys-Thr-NH₂;

(H)(3-[p-hydroxyphenyl])-D-Cys-Tyr-D-Trp-Lys-Val-Cys-β-Nal-NH₂;

(H)(3-naphthyl]propionyl)-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-β-Nal-NH₂;

(H)(3-naphthyl]propionyl)-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-Thr-NH₂;

(H)(3-phenylylpropionyl)-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-β-Nal-NH₂;

(H)(3-phenylylpropionyl)-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-Thr-NH₂;

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(CH₃CO)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(CH₃CO)-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(CH₃CO)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(CH₃CO)-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

H₂-Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(CH₃CO)Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

H₂-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

H(CH₃CO)Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

H₂-Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(CH₃CO)Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

H₂-Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(CH₃CO)Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

(H)(CH₃CO)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

(H)(CH₃CO)-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl) ethyl amide;

(H)(CH₂CO)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl)ethylamide;

H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl)ethylamide;

(H)(CH₃CO)-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl)ethylamide;

H₂-Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

(H)(CH₃CO)Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)Phe-D-Cys-Tyr-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

H₂-Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R-(2-naphthyl) ethylamide;

(H)(CH₃CO)Phe-Cys-Pal-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)Phe-D-Cys-Pal-D-Trp-Lys-Val-Cys-2R-(2-naphthyl)ethylamide;

H₂-Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl) ethylamide;

(H)(CH₃CO)Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)Phe-D-Cys-Tyr-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl)ethylamide;

H₂-Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl)ethylamide;

(H)(CH₃CO)Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl)ethylamide;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)Phe-D-Cys-Pal-D-Trp-Lys-Thr-Cys-2R-(2-naphthyl)ethylamide;

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-2R-(2-naphthyl)ethylamide;

H₂-Phe-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-2R-(2-naphthyl)ethylamide;

H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

H₂-Phe-D-Cys-Tyr-D-Trp-Lys-Abu-Cys-2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide;

H₂-Phe-D-Phe-Tyr-D-Trp-Lys-Thr-Phe-Thr-NH₂;

H₂-Phe-D-Phe-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂;

H₂-Phe-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂;

H₂-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂ (Analog No. 1);

(H)(CH₃CO)-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂;

H₂-β-Nal-D-Cpa-Pal-D-Trp-Lys-Val-Phe-Thr-NH₂;

(H)(CH₃CO)-β-Nal-D-Cpa-Pal-D-Trp-Lys-Val-Phe-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cpa-Pal-D-Trp-Lys-Val-Phe-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cpa-Pal-D-Trp-Lys-Val-Phe-Thr-NH₂;

H₂-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Thr-Phe-Thr-NH₂;

(H)(CH₃CO)-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Thr-Phe-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Thr-Phe-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Thr-Phe-Thr-NH₂;

H₂-β-Nal-D-Cpa-Pal-D-Trp-Lys-Thr-Phe-Thr-NH₂;

(H)(CH₃CO)-β-Nal-D-Cpa-Pal-D-Trp-Lys-Thr-Phe-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cpa-Pal-D-Trp-Lys-Thr-Phe-Thr-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cpa-Pal-D-Trp-Lys-Thr-Phe-Thr-NH₂;

H₂-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-β-Nal-NH₂;

(H)(CH₃CO)-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-β-Nal-NH₂;

(H)(4-(2-hydroxyethyl)-1-piperazinylacetyl)-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-β-Nal-NH₂;or

(H)(4-(2-hydroxyethyl)-1-piperizineethanesulfonyl)-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-β-Nal-NH₂;

H₂-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-β-Nal-NH₂ (Analog No. 23);

H₂-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂;

H₂-D-β-Nal-D-Cpa-Phe-D-Trp-Lys-Val-Phe-Thr-NH₂;

H₂-D-β-Nal-D-Phe-Tyr-D-Trp-Lys-Thr-Phe-Thr-NH₂;

H₂-D-Phe-D-Phe-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂;

H₂-D-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂ (Analog No. 8); or

H₂-D-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-β-Nal-NH₂; or

a pharmaceutically acceptable salt thereof.

With the exception of the N-terminal amino acid, all abbreviations(e.g., Ala or A₂) of amino acids in this disclosure stand for thestructure of —NH—CH(R)—CO—, wherein R is a side chain of an amino acid(e.g., CH₃ for Ala). For the N-terminal amino acid, the abbreviationstands for the structure of ═N—CH(R)—CO—, wherein R is a side chain ofan amino acid. Pen, β-Ala, Gaba, Nle, Nva, Pal, F₅-Phe,2,4-dichloro-Phe, Cpa, β-Nal, β-1-Nal, Abu, Dip, 2-Pal, Bip, Npa, Igl,Bta, Tle, Bpa, Iph, Cha, Thr(Bzl), Tyr(Bzl), and Aib are respectiveabbreviations of the following α-amino acids: penicillamine,3-aminopropionic acid, 4-aminobutyric acid, norleucine, norvaline,β-[3-pyridyl]-alanine, [2,3,4,5,6-pentafluorophenyl]-alanine,β-[2,4-dichlorophenyl]-alanine, β-[4-chlorophenyl]-alanine,β-[2-napthyl]-alanine, β-[1-naphthyl]-alanine; 2-aminobutyric acid,3,3′-diphenylalanine, [2-pyridyl]-alanine, 4,4′-biphenylalanine,p-NO₂-phenylalanine, 0.2-indanylglycine, 3-benzothienylalanine,α-[t-butyl]-glycine, 4-bromo-phenylalanine, 4′-iodo-phenylalanine,β-(cyclohexyl)-alanine, O-benzyl-threonine, O-benzyl-tyrosine, and2-aminoisobutyric acid. Tyr(I) refers to an iodinated tyrosine residue(e.g., 3-I-Tyr, 5-I-Tyr, 3,5-I-Tyr) wherein the iodine may be aradioactive isotope, e.g., I¹²⁵, I¹²⁷, or I¹³¹. An aliphatic amino acidis an α-amino acid having one or two side chains which, independently,are hydrocarbons, e.g., a straight or branched chain of 1-6 carbons.Examples of aliphatic amino acids include Ala, Aib, Val, Leu, Tle, Ile,Nle, Nva, or Abu. An aromatic amino acid is an α-amino acid the sidechain of which has a neutral (e.g., not acidic or basic) aromaticsubstituent, e.g., a substituted or unsubstituted phenyl, naphthyl, oraromatic heterocycle group (e.g., pyridyl or indolyl). Examples ofaromatic amino acids include Phe, p-X-Phe (where X is a halo (e.g., F,Cl, Br, or I), OH, OCH₃, CH₃, or NO₂), o-X-Phe (where X is a halo, OH,OCH₃, CH₃, or NO₂), m-X-Phe (where X is a halo, OH, OCH₃, CH₃, or NO₂),His, Pal, Trp, β-Nal, 2,4-dichloro-Phe, Tyr(I),β-[3,4,5-trifluorophenyl]-alanine, Bta, β-[3-cyanophenyl]-alanine,β-[4-cyanophenyl]-alanine, β-[3′,4-difluorophenyl]-alanine,β-[3,5-difluorophenyl]-alanine, β-[2-fluorophenyl]-alanine,β-[4-thiazolyl]-alanine, Bip, Dip, Npa, Igl, Bpa, Iph,homophenylalanine, 2-Pal, β-[4-pyridyl]-alanine,β-[4-thiazolyl]-alanine, β-[2-thiazolyl)-alanine,para-(CF₃)-phenylalanine, and F₅-Phe. What is meant by an “Eaa” is anamino acid of the formula —NH—[CH(R)_(n)—CO— (where n is 2-6 and R is H,lower alkyl, or hydroxy lower alkyl). Examples of an Eaa include β-Alaand Gaba.

As used herein, “lower alkyl” is intended to include both branched andstraight-chain saturated aliphatic hydrocarbon groups having 1-6 carbonatoms. Examples of lower alkyl groups include methyl, ethyl, propyl,isopropyl, butyl, t-butyl, isobutyl, sec-butyl, and the like.

As used herein, “aryl”, is intended to include any stable monocyclic,bicyclic, or tricyclic carbon ring(s) of up to 7 members in each ring,wherein at least one ring is aromatic. Examples of aryl groups includephenyl, naphthyl, anthracenyl, biphenyl, tetrahydronaphthyl, indanyl,phenanthrenyl, and the like.

The term “heterocyclyl”, as used herein, represents a stable 5- to7-membered monocyclic or stable 8- to 11-membered bicyclic or stable11-15 membered tricyclic heterocyclic ring which is either saturated orunsaturated, and which consists of carbon atoms and from one to fourheteroatoms selected from the group consisting of N, O, and S, andincluding any bicyclic group in which any of the above-definedheterocyclic rings is fused to a benzene ring. The heterocyclic ring maybe attached at any heteroatom or carbon atom which results in thecreation of a stable structure. Examples of such heterocyclic elementsinclude, but are not limited to, azepinyl, benzimidazolyl,benzisoxazolyl, benzofurazanyl, benzopyranyl, benzothiopyranyl,benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, chromanyl,cinnolinyl, dihydrobenzofuryl, dihydrobenzothienyl,dihydrobenzothiopyranyl, dihydrobenzothio-pyranyl sulfone, furyl,imidazolidinyl, imidazolinyl, imidazolyl, indolinyl, indolyl,isochromanyl, isoindolinyl, isoquinolinyl, isothiazolidinyl,isothiazolyl, isothiazolidinyl, morpholinyl, naphthyridinyl,oxadiazolyl, 2-oxoazepinyl, 2-oxopiperazinyl, 2-oxopiperidinyl,2-oxopyrrolidinyl, piperidyl, piperazinyl, pyridyl, pyridyl N-oxide,quinoxalinyl, tetrahydrofuryl, tetrahydroisoquinolinyl,tetrahydro-quinolinyl, thiamorpholinyl, thiamorpholinyl sulfoxide,thiazolyl, thiazolinyl, thienofuryl, thienothienyl, thienyl, and thelike.

The term “substituted” is meant to include the recited chemical group(e.g., lower alkyl, heterocycle, aryl, cycloalkyl, etc.) substitutedwith one to four of the recited substituents (e.g., halo, hydroxy, loweralkyl, etc.). The substituent may be attached to any atom in thechemical group.

The structure of 4-(2-hydroxyethyl)]-1-piperazinylacetyl and4-(2-hydroxyethyl)]-1-piperizineethanesulfonyl are, respectively, asfollows:

The compounds of this invention can be provided in the form ofpharmaceutically acceptable salts. Acceptable salts include, but are notlimited to, acid addition salts of inorganic acids such ashydrochloride, sulfate, phosphate, diphosphate, hydrobromide, andnitrate or organic acids such as acetate, maleate, fumarate, tartrate,succinate, citrate, lactate, methanesulfonate, p-toluenesulfonate,pamoate, salicylate, oxalate, and stearate. Also within the scope of thepresent invention, where applicable, are salts formed from bases such assodium or potassium hydroxide. For further examples of pharmaceuticallyacceptable salts see, “Pharmaceutical Salts,” J. Pharm. Sci. 66:1(1977).

Where the amino acid residue is optically active, it is the L-isomerthat is intended unless otherwise specified. In the formulae set forthherein, the disulfide bond between the thiol group on the side chain ofresidue A₂ (e.g., Cys, Pen, D-Cys, or D-Pen) and the thiol group on theside chain of residue A₇ (e.g., Cys or Pen) is not shown.

The peptides of the invention can be used to promote the release ofgrowth hormone or insulin in a subject (e.g., a mammal such as a humanpatient). Thus, the peptides are useful in the treatment ofphysiological conditions in which the promotion of the release of growthhormone or insulin is of benefit. The peptides of the invention can alsobe used in enhancing wound healing or promoting angiogenesis. Also,peptides of the invention having a Tyr(I) residue can be used to imagecells containing somatostatin receptors. Such peptides of the inventioncan be used either in vivo to detect cells having somatostatin receptors(e.g., cancer cells) or in vitro as a radioligand in a somatostatinreceptor binding assay. The peptide of the invention can also be used asvectors to target cells with radioactive isotopes.

A therapeutically effective amount of a peptide of this invention and apharmaceutically acceptable carrier substance (e.g., magnesiumcarbonate, lactose, or a phospholipid with which the therapeuticcompound can form a micelle) together form a therapeutic composition(e.g., a pill, tablet, capsule, or liquid) for administration (e.g.,orally, intravenously, transdermally, pulmonarily, vaginally,subcutaneously, nasally, iontophoretically, or by intratracheally) to asubject in need of the peptide. The pill, tablet, or capsule can becoated with a substance capable of protecting the composition from thegastric acid or intestinal enzymes in the subject's stomach for a periodof time sufficient to allow the composition to pass undigested into thesubject's small intestine. The therapeutic composition can also be inthe form of a biodegradable or nonbiodegradable sustained releaseformulation for subcutaneous or intramuscular administration. See, e.g.,U.S. Pat. Nos. 3,773,919 and 4,767,628 and PCT Application No. WO94/00148. Continuous administration can also be obtained using animplantable or external pump (e.g., INFUSAID™ pump) to administer thetherapeutic composition.

The dose of a peptide of the present invention for treating theabove-mentioned diseases or disorders varies depending upon the mannerof administration, the age and the body weight of the subject, and thecondition of the subject to be treated, and ultimately will be decidedby the attending physician or veterinarian. Such an amount of thepeptide as determined by the attending physician or veterinarian isreferred to herein as a “therapeutically effective amount.”

Also contemplated within the scope of this invention is a peptidecovered by the above generic formula for both use in treating diseasesor disorders associated with the need to promote the release of growthhormone or insulin, and use in detecting somatostatin receptors, e.g.,radioimaging.

Other features and advantages of the present invention will be apparentfrom the detailed description and from the claims.

DETAILED DESCRIPTION OF THE INVENTION

It is believed that one skilled in the art can, based on the descriptionherein, utilize the present invention to its fullest extent. Thefollowing specific embodiments are, therefore, to be construed as merelyillustrative, and not limitative of the remainder of the disclosure inany way whatsoever.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Also, all publications, patentapplications, patents, and other references mentioned herein areincorporated by reference.

Synthesis

The synthesis of short peptides is well examined in the peptide art. Seee.g., Stewart, et al., Solid Phase Peptide Synthesis (Pierce ChemicalCo., 2d ed., 1984). The following describes the synthesis ofD-β-Nal-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂ andD-β-Nal-Cpa-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂. Other peptides of theinvention can be prepared in an analogous manner by a person of ordinaryskill in the art.

(a) Synthesis of H₂-β-Nal-D-Cys-Pal-D-Trp-Lys-Val-Cys-β-Nal-NH₂

-   -   1)        Boc-β-naphthylalanine-S-methylbenzyl-D-cysteine-3-pyridyl-2-alanine-D-tryptophan-N^(e)-benzyloxycarbonyl-lysine-valine-S-methylbenzyl-cysteine-β-naphthylalanine-benzhydrylamine        resin.

Benzhhydrylamine-polystyrene resin (Advanced ChemTech, Inc., Louisville,Ky.) (1.2 g; 0.5 mmole) in the chloride ion form was placed in thereaction vessel of an Advanced ChemTech™ peptide synthesizer programmedto perform the following reaction cycle: (a) methylene chloride; (b) 33%trifluoroacetic acid in methylene chloride (2 times for 1 and 25 mineach); (c) methylene chloride; (d) ethanol; (e) methylene chloride; and(f) 10% triethylamine in chloroform.

The neutralized resin was stirred with Boc-O-β-naphthylalanine anddiisopropylcarbodiimide (1.5 mmole each) in methylene chloride for 1 hr,and the resulting amino acid resin was then cycled through steps (a) to(f) in the above wash program. The following amino acids (1.5 mmole)were then coupled successively by the same procedure:.Boc-S-methylbenzyl-Cys, Boc-Val, Boc-N^(e)-benzyloxycarbonyl-lysine,Boc-D-Trp, Boc-Pal, and Boc-S-methylbenzyl-D-Cys and Boc-β-Nal. Afterwashing and drying, the completed resin weighed 2.0 g.

-   -   2)        β-naphthylalanine-c[D-cysteine-3-pyridyl-2-alanine-D-tryptophan-lysine-valine-cysteine]-β-naphthylalanine-NH₂

The completed resin described in (1) (1.0 g, 0.25 mmole) was mixed withanisole (5 ml), dithiothreitol (100 mg), and anhydrous hydrogen fluoride(35 ml) at 0° C. and stirred for 45 min. Excess hydrogen fluoride wasevaporated rapidly under a stream of dry nitrogen, and the free peptideis precipitated and washed with ether. The crude peptide was thendissolved in 500 ml of 90* acetic acid to which was added a concentratedsolution of I₂/MeOH until a permanent brown color is observed. Excess I₂is removed by addition of ascorbic acid and the solution evaporated to asmall volume which was applied to a column (2.5×90 cm) of Sephadex™G-25, which was eluted with 50 t AcOH. Fractions containing a majorcomponent by ultraviolet (UV) absorption and thin layer chromatographywere then pooled, evaporated to a small volume, and applied to a column(1.5×70 cm) of Vydac™ octadecylsilane silica (10-15 μm). This was elutedwith a linear gradient of acetonitrile in 0.1% trifluoroacetic acid inwater. Fractions were examined by thin layer chromatography (TLC) andanalytical high performance liquid chromatography (HPLC) and pooled togive maximum purity. Repeated lyophilization of the solution from watergave the desired product as a white, fluffy powder. The product wasfound to be homogeneous by HPLC and TLC. Amino acid analysis of an acidhydrolysate and matrix-assisted laser desorption (MALD) massspectroscopy confirmed the composition of the octapeptide.

(b) Synthesis of H₂-D-β-Nal-Cpa-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂

1)Boc-β-D-naphthylalanine-D-4-chlorophenylalanine-O-dichlorobenzyl-tyrosine-D-tryptophan-N^(e)-benzyloxycarbonyl-lysine-valine-S-phenylalanine-O-benzyl-threonine-benzhydrylamineresin

Benzhydrylamine-polystyrene resin (Advanced ChemTech™, Inc.) (1.2 g, 0.5mmole) in the chloride ion form was placed in the reaction vessel of anAdvanced ChemTech peptide synthesizer programmed to perform thefollowing reaction cycle: (a) methylene chloride; (b) 33%trifluoroacetic acid in methylene chloride (2 times for 1 and 25 mineach); (c) methylene chloride; (d) ethanol; (e) methylene chloride; and(f) 10% triethylamine in chloroform.

The neutralized resin was stirred with Boc-O-benzylthreonine anddiisopropylcarbodiimide (1.5 mmole each) in methylene chloride for 1 hrand the resulting amino acid resin was cycled through steps (a) to. (f)in the above wash program. The following amino acids (1.5 mmole) werethen coupled successively by the same procedure: Boc-phenylalanine,Boc-Val, Boc-N^(e)-benzyloxycarbonyl-lysine, Boc-D-Trp,Boc-O-dichlorobenzyl-Tyr, and Boc-D-4-chlorophenylalanine, andBoc-β-D-Nal. After washing and drying, the completed resin weighed 2.1g.

2)β-D-naphthylalanine-D-4-chlorophenylalanine-tyrosine-D-tryptophan-lysine-valine-phenylalanine-threonine-NH₂

The peptide resin from (1) was subjected to HF cleavage as describedabove. Column purification as described yielded the desired compound asa white, fluffy powder (170 mg) which is found to be homogeneous by HPLCand TLC. Amino acid analysis of an acid hydrolysate and MALD massspectroscopy confirms the composition of the peptide.

Peptides containing C-terminal substituted amides can be made by solidphase methods by displacing the appropriate peptide off the solid phasewith the corresponding amine. Alternatively, these analogs may besynthesized by solution-phase peptide synthesis methods in which thegrowing peptide chain is maintained in solution in an organic solventduring synthesis and assembled by iterative coupling/deprotectioncycles. Final removal of the side chain protecting groups yields thedesired peptide after appropriate purification. Peptides containingN-terminal substitutions (e.g., where R₁ is E, CO, or E₁SO₂ (where E₁ isheterocycle lower alkyl) substituted with hydroxy lower alkyl and R₂ isH such as 4-(2-hydroxyethyl)-1-piperazinylacetyl or4-(2-hydroxyethyl)-1-piperdineethanesulfonyl) can be synthesized asdescribed in PCT Application No. WO 95/04752.

Bioassay on the In Vitro Release of Growth Hormone

(a) Rat Pituitary Cell Dispersion

Pituitaries from adult Charles River CD male rats (Wilmington, Mass.)housed under controlled conditions were dispersed and cultured usingaseptic technique by modification of previously described methods(Hoefer, M. T., et al., Mol. Cell. Endocrinol. 35:229 (1984);Ben-Jonathan, N., et al., Methods Enzymol. 103:249 (1983); and Heiman,M. L., et al., Endocrinology 116:410 (1985)). Pituitaries were removedfrom sacrificed rats, sectioned, and then placed into a siliconized,liquid scintillation vial containing 2 ml 0.2% trypsin (WorthingtonBiochemicals, Freehold, N.J.) in sterile-filtered Krebs-Ringerbicarbonate buffer supplemented with 1% bovine serum albumin, 14 mMglucose, modified Eagle medium (MEM) vitamin solution, and MEM aminoacids (Gibco Laboratories, Grand Island, N.Y.) (KRBGA). All glasswarewas siliconized as described by Sayers, et al., Endocrinology 88:1063(1971). The fragments were incubated in a water bath for 35 min at 37°C. with agitation. The vial contents then were poured into ascintillation vial containing 2 ml 0.1% DNase (Sigma Chemical Co., St.Louis, Mo.) in KRBGA and incubated for 2 min at 37° C. with agitation.After incubation, the tissue was decanted into a 15 ml centrifuge tubeand allowed to settle. Medium was discarded, and-pituitary sections werewashed 3 times with 1 ml fresh KRBGA. The cells were then dispersed in 2ml 0.05% LBI (lima bean trypsin inhibitor, Worthington Biochemicals) bygently drawing the fragments into and expelling them out of asiliconized, fire polished Pasteur pipette. Dispersed cells were thenfiltered through a 630 μm diameter Nylon mesh (Tetko, Elmsford, N.Y.)into a fresh 15 ml centrifuge tube. An additional 2 ml of 0.05% LBIsolution was used to rinse the first tube and was transferred to thesecond tube with filtering.

(b) Cell Culture

The dispersed cells were then further diluted with approximately 15 mlsterile-filtered Dulbecco's modified Eagle medium (GIBCO), which wassupplemented with 2.5% fetal calf serum (GIBCO), 3% horse serum (GIBCO),10% fresh rat serum (stored on ice for no longer than 1 hr) from thepituitary donors, 1% MEM non-essential amino acids (GIBCO), andgentamycin (10 ng/ml; Sigma) and nystatin (10,000 U/ml; GIBCO). Thecells were poured into a 50 ml round-bottomed glass extraction flaskwith a large diameter opening and then randomly plated at a density ofapproximately 200,000, cells per well (Co-star cluster 24; RochesterScientific Co., Rochester, N.Y.). The plated cells were maintained inthe above Dulbecco's medium in a humidified atmosphere of 95% air and 5%CO, at 37° C. for 4-5 days.

(c) Experimental Incubation and IC₅₀ Determination

In preparation for a hormone challenge, the cells were washed 3 timeswith medium 199 (GIBCO) to remove old medium and floating cells. Eachtreatment well contained a total volume of 1 ml medium 199 containing 1%BSA (fraction V; Sigma) with treatments as described below. Eachantagonist candidate was tested using a single 24-well cell cultureplate. Each treatment was performed in triplicate. Each plate contained8 treatment groups: one 1 nM growth hormone releasing factor (GRF)(1-29)NH₂-stimulated control group; one 1 nM somatostatin-inhibitedcontrol group in the presence of 1 nM GRF(1-29)NH₂; and 6 doses of agiven antagonist in the presence of both 1 nM SRIF and 1 nM GRF perplate. After 3 hrs at 37° C. in a air/carbon dioxide atmosphere (95/5%),the medium was removed and stored at −20° C. until radioimmunoassayedfor growth hormone content. IC₅₀'s of each antagonist versus 1 nM SRIFwere calculated using a computer program (SigmaPlot, Jandel Scientific,San Rafael, Calif.) with the maximum response constrained to the valueof the 1 nM GRF(1-29)NH₂-stimulated control. These IC₅₀'s, are presentedin Table I.

TABLE I ANALOG NO. IC₅₀ (μM) 1 3.03 2 0.04 3 0.01 4 0.03 5 0.06 6 0.9 70.071 8 3.96 9 1.36 10 0.62 11 0.72 12 0.056 13 0.11 14 0.11 15 0.14 160.82 17 1 18 0.38 19 0.11 20 0.12 21 0.97 22 0.066 23 0.91 24 0.068 250.28 26 0.38 27 0.041 28 0.10 29 0.0084 30 0.0065 31 0.0038 32 0.012 331.50 34 0.42 35 0.052 36 1.03 37 0.78 38 0.11 39 0.034 40 0.11 41 0.2142 0.044 43 0.00082 44 0.021 45 0.13 46 0.02 47 0.053 48 0.050 49 0.2350 0.0011 51 0.012 52 0.0026 53 0.0029 54 0.029 55 0.0026 56 0.0018 570.0059

Other Embodiments

It is to be understood that while the invention has been described inconjunction with the detailed description thereof, that the foregoingdescription is intended to illustrate and not limit the scope of theinvention, which is defined by the scope of the appended claims. Otheraspects, advantages, and modifications are within the claims.

1. A compound of the formula: H₂-β-Nal-D-Cys-Tyr-D-Trp-Lys-Val-Cys-(2R,3R-(2-hydroxymethyl)-3-hydroxy)propylamide; H₂-Phe-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂; H₂-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂; H₂-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-β-Nal-NH₂; H₂-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂; H₂-D-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-Thr-NH₂; H₂-D-β-Nal-D-Cpa-Tyr-D-Trp-Lys-Val-Phe-β-Nal-NH₂;or H₂-Phe-D-Phe-Tyr-D-Trp-Lys-Thr-Phe-Thr-NH₂; or a pharmaceutically acceptable salt thereof. 